Aspergillus Mycotoxin Research Articles

Evaluation Of Some Aspergillus Mycotoxins In Patients With Bronchitis In Al-Najaf City

This research aimed to establish whether the Aspergillus species identified from bronchitis patients were mycotoxigenic or non-mycotoxigenic fungi in sputum and serum samples. A total sample of 170 subjects with ages ranging from 15 to 72 years— were obtained from Al-Najaf Province (Al-Sadr Medical City) between December 2023 and April 2024, divided into patient (n=176) and healthy groups (n=34). The Aspergillus genera isolated were cultured on coconut agar medium and screened under UV light (360 nm); each sputum and serum sample was also tested using the ELISA technique. All Aspergillus strains exhibited blue fluorescence on Coconut Cream Agar (CCA) media under UV light, confirming positivity. The study identified production of aflatoxins B1 and ochratoxins-A in sputum and serum samples among both patients with bronchitis and control groups. The results showed a significant difference in patient groups by samples (P≤ 0.05) while the healthy groups showed a non-significant (P>0.05) difference. Most Aspergillus spp. Isolated from bronchitis patients were found to produce aflatoxin B1 and ochratoxin-A on coconut agar medium— mycotoxins extracts included sputum (about 68.4% for aflatoxins-B1 and 41.7% for serum samples) and sputum (about 77.6% for ochratoxin-A and 91.7% for serum samples) taken from bronchitis patients, which were identified using ELISA technique.

Endocrine Effect of Some Mycotoxins on Humans: A Clinical Review of the Ways to Mitigate the Action of Mycotoxins.

Fungi such as Aspergillus spp. and Fusarium spp., which are commonly found in the environment, pose a serious global health problem. This study aims to present the results of epidemiological studies, including clinical cases, on the relationship between human exposure to some mycotoxins, especially zearalenone and aflatoxin, and the occurrence of reproductive disorders. In addition, examples of methods to reduce human exposure to mycotoxins are presented. In March 2023, various databases (PubMed, Google Scholar, EMBASE and Web of Science) were systematically searched using Google Chrome to identify studies evaluating the association between exposure to mycotoxins and the occurrence of complications related to impaired fertility or cancer incidence. The analysed data indicate that exposure to the evaluated mycotoxins is widespread and correlates strongly with precocious puberty, reduced fertility and increased cancer incidence in women and men worldwide. There is evidence to suggest that exposure to the Aspergillus mycotoxin aflatoxin (AF) during pregnancy can impair intrauterine foetal growth, promote neonatal jaundice and cause perinatal death and preterm birth. In contrast, exposure to the Fusarium mycotoxin zearalenone (ZEA) leads to precocious sexual development, infertility, the development of malformations and the development of breast cancer. Unfortunately, the development of methods (biological, chemical or physical) to completely eliminate exposure to mycotoxins has limited practical application. The threat to human health from mycotoxins is real and further research is needed to improve our knowledge and specific public health interventions.

New perspectives in application of kidney biomarkers in mycotoxin induced nephrotoxicity, with a particular focus on domestic pigs.

The gradual spread of Aspergilli worldwide is adding to the global shortage of food and is affecting its safe consumption. Aspergillus-derived mycotoxins, including aflatoxins and ochratoxin A, and fumonisins (members of the fusariotoxin group) can cause pathological damage to vital organs, including the kidney or liver. Although the kidney functions as the major excretory system in mammals, monitoring and screening for mycotoxin induced nephrotoxicity is only now a developmental area in the field of livestock feed toxicology. Currently the assessment of individual exposure to mycotoxins in man and animals is usually based on the analysis of toxin and/or metabolite contamination in the blood or urine. However, this requires selective and sensitive analytical methods (e.g., HPLC-MS/MS), which are time consuming and expensive. The toxicokinetic of mycotoxin metabolites is becoming better understood. Several kidney biomarkers are used successfully in drug development, however cost-efficient, and reliable kidney biomarkers are urgently needed for monitoring farm animals for early signs of kidney disease. β2-microglobulin (β2-MG) and N-acetyl-β-D-glucosaminidase (NAG) are the dominant biomarkers employed routinely in environmental toxicology research, while kidney injury molecule 1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL) are also emerging as effective markers to identify mycotoxin induced nephropathy. Pigs are exposed to mycotoxins due to their cereal-based diet and are particularly susceptible to Aspergillus mycotoxins. In addition to commonly used diagnostic markers for nephrotoxicity including plasma creatinine, NAG, KIM-1 and NGAL can be used in pigs. In this review, the currently available techniques are summarized, which are used for screening mycotoxin induced nephrotoxicity in farm animals. Possible approaches are considered, which could be used to detect mycotoxin induced nephropathy.

The Toll pathway mediates Drosophila resilience to Aspergillus mycotoxins through specific Bomanins

Host defense against infections encompasses both resistance, which targets microorganisms for neutralization or elimination, and resilience/disease tolerance, which allows the host to withstand/tolerate pathogens and repair damages. In Drosophila, the Toll signaling pathway is thought to mediate resistance against fungal infections by regulating the secretion of antimicrobial peptides, potentially including Bomanins. We find that Aspergillus fumigatus kills Drosophila Toll pathway mutants without invasion because its dissemination is blocked by melanization, suggesting a role for Toll in host defense distinct from resistance. We report that mutants affecting the Toll pathway or the 55C Bomanin locus are susceptible to the injection of two Aspergillus mycotoxins, restrictocin and verruculogen. The vulnerability of 55C deletion mutants to these mycotoxins is rescued by the overexpression of Bomanins specific to each challenge. Mechanistically, flies in which BomS6 is expressed in the nervous system exhibit an enhanced recovery from the tremors induced by injected verruculogen and display improved survival. Thus, innate immunity also protects the host against the action of microbial toxins through secreted peptides and thereby increases its resilience to infection.

Changes in Fusarium and Aspergillus Mycotoxin Content and Fatty Acid Composition after the Application of Ozone in Different Maize Hybrids.

Mycotoxins in maize represent a great threat to human health. For this reason, novel technics such as ozone treatment are used to reduce the content of maize mycotoxins. However, there is little knowledge about the effect of ozone treatment on maize quality parameters. This study investigated the changes in Fusarium and Aspergillus mycotoxins and the changes in fatty acids during the ozone treatment of maize samples. Sixteen maize hybrids were visually tested for the naturally occurring ear rot severity and treated with three different concentrations of ozone (40, 70, and 85 mg/L). Mycotoxin content in maize samples was determined using a high-performance liquid chromatography (HPLC) system, whereas dominant fatty acids were determined using gas chromatography coupled with a flame ionization detector (GC-FID). Ozone treatments could be successfully applied to reduce the content of mycotoxins in maize below the detection limit. Ozone treatments increased the content of monounsaturated fatty acids (MUFAs) and decreased the content of polyunsaturated fatty acids (PUFAs), i.e., linoleic acid (36.7% in relation to the lowest applied ozone concentration), which negatively affected the nutritional value of maize.

The Most Important Secondary Metabolites of the Genus Aspergillus

The presence of Aspergillus mycotoxins has been confirmed in various feed and food. They represent a hidden threat that cannot be completely prevented. In animal husbandry, they impair their productive health and, in human nutrition, they can cause acute or chronic health complications. This article provides an overview of the most important secondary metabolites of the genus Aspergillus in terms of their occurrence, toxicity and physico-chemical properties.

Conventional vs. organic vineyards: Black Aspergilli population structure, mycotoxigenic capacity and mycotoxin contamination assessment in wines, using a new Q-TOF MS-MS detection method

Aspergillus bunch rot, caused by several mycotoxigenic species of Black Aspergilli section Nigri is one of the most severe diseases of grapevine, while mycotoxin contaminated products may pose a risk to the consumers health. During a 2-year (2018–2019) survey, the impact of conventional and organic farming systems on Black Aspergilli population structure in Greek vineyards was thoroughly investigated. In total, 300 isolates of Aspergillus spp. were selected and identified by amplicon sequencing of the ITS-region, β-tubulin and calmodulin genes. Four different Aspergillus section Nigri species (A. tubingensis, A. uvarum, A. carbonarius and A. niger) were identified as the casual agents of the disease. During both sampling years, higher frequencies of A. tubingensis and A. uvarum were found in the organic and conventional vineyards, respectively. In vitro production of ochratoxin A (OTA) and fumonisin B2, B3 and B4 (FB2, FB3, and FB4) was evaluated in two selective media. The analysis revealed a low frequency of mycotoxigenic isolates, while, most of these isolates originated from conventional vineyards. As expected, A. carbonarious was identified as the main OTA producer (0.01–9.2 μg/g), whereas A. niger was the leading producer of FBs. In total, 74 wine samples originating from either organic or conventional vineyards of the same region, were analyzed using a new Quadrupole Time-of-Flight Mass Spectrometry (QTOF MS-MS) analytical method developed to detect and quantify Alternariol (AOH), Alternariol monomethyl ether (AME), and Citrinin (CIT) in addition to the aforementioned Aspergillus mycotoxins (OTA, FB1, FB2 FB3), using a modified QuEChERS extraction protocol. Interestingly, a low frequency of mycotoxin-contaminated wines was detected. At the same time, fumonisins (FBs) were identified as the main mycotoxins in Greek wines. This is the first study deciphering the impact of conventional and organic farming systems on Aspergillus section Nigri species in Greek vineyards, suggesting that farming system may affect the species composition within the vineyard. Furthermore, evidence was provided for a higher risk of selection of Aspergillus strains with higher mycotoxigenic capacity in conventional vineyards, associated with higher risk of mycotoxin contamination in wines originating from these vineyards.

Biodiversity and Distribution of <i>Aspergillus</i> and Their Toxins in Maize from Western and Eastern Regions of South Africa

Aspergillus species and aflatoxins production are more prevalent during times of high heat and drought. In South Africa, there is frequent occurrence of drought as a result of climate change. The aim of this study was to investigate the biodiversity and distribution of Aspergillus species with their corresponding toxins in maize from main maize producing regions of South Africa; [Western Regions (WR) and Eastern Regions (ER)]. One hundred and twenty-three (64 from WR and 59 from ER) maize samples from the two agro-climatic regions in South Africa were analyzed using cultural, molecular and analytical methods. Across agro-climatic regions, Aspergillus species contaminated about 62% of the maize samples, while Aspergillus flavus was the most prevalent (47.15%) followed by Aspergillus fumigatus (4.69%) while the least was Aspergillus parasiticus (0.81%). The Western Regions showed a higher distribution of varieties of Aspergillus species compared to the Eastern Regions. Aflatoxins contaminated only 27.64% of the maize samples with a mean total aflatoxin concentration of 2.40 μg/kg which is below the South Africa’s set standard for total aflatoxin in food (5 μg/kg). About 10.57% of the samples produce aflatoxins above the 5 μg/kg permissible limit for total aflatoxin in foods. The ratio of toxigenic to atoxigenic strains of Aspergillus flavus was generally low in all the regions of South Africa. This study could aid policy makers to make informed decisions in developing remediation strategies for Aspergillus mycotoxins.

Degradation of Aflatoxins B1 by Atoxigenic Aspergillus flavus Biocontrol Agents.

Aflatoxins are potent Aspergillus mycotoxins that contaminate food and feed, thereby impacting health and trade. Biopesticides with atoxigenic Aspergillus flavus isolates as active ingredients are used to reduce aflatoxin contamination in crops. The mechanism of aflatoxin biocontrol is primarily attributed to competitive exclusion but, sometimes, aflatoxin is reduced by greater amounts than can be explained by displacement of aflatoxin-producing fungi on the crop. Objectives of this study were to (i) evaluate the ability of atoxigenic A. flavus genotypes to degrade aflatoxin B1 (AFB1) and (ii) characterize impacts of temperature, time, and nutrient availability on AFB1 degradation by atoxigenic A. flavus. Aflatoxin-contaminated maize was inoculated with atoxigenic isolates in three separate experiments that included different atoxigenic genotypes, temperature, and time as variables. Atoxigenic genotypes varied in aflatoxin degradation but all degraded AFB1 >44% after 7 days at 30°C. The optimum temperature for AFB1 degradation was 25 to 30°C, which is similar to the optimum range for AFB1 production. In a time-course experiment, atoxigenics degraded 40% of AFB1 within 3 days, and 80% of aflatoxin was degraded by day 21. Atoxigenic isolates were able to degrade and utilize AFB1 as a sole carbon source in a chemically defined medium but quantities of AFB1 degraded declined as glucose concentrations increased. Degradation may be an additional mechanism through which atoxigenic A. flavus biocontrol products reduce aflatoxin contamination pre- or postharvest. Thus, selection of optimal atoxigenic active ingredients can include assessment of both competitive ability in agricultural fields and their ability to degrade aflatoxins.

Antifungal activity and the effect of the essential oil of Lippia origanoides Kunth on Aspergillus mycotoxins production

Lippia origanoides (Verbenaceae) is an aromatic plant of great importance in Brazilian medicine. The antifungal and antimycotoxigenic effects of Lippia origanoides essential oil (LEO) on Aspergillus carbonarius, Aspergillus flavus and Aspergillus ochraceus were investigated. The LEO was obtained by hydrodistillation, and it was characterized by GC-FID and GC-MS. The antifungal activity was determined by mycelial growth and mycotoxin and ergosterol assays. Scanning electron microscopy was used to evaluate the effect of the LEO on the plasma membrane of fungi. The main constituents in the LEO were carvacrol (58.72%), ρ-cymene (15.37%) and γ-terpinene (12.73%). Significant fungicidal effects of LEO against A. carbonarius (0.24 μL/mL), A. flavus (0.98 μL/mL) and A. ochraceus (0.98 μL/mL) were observed. A reduction in the ochratoxin A synthesis that ranged from 92.03 to 98.02% and from 81.72 to 96.92% for the fungal species A. carbonarius and A. ochraceus, respectively, was observed in the presence of LEO. The production of aflatoxin B1 and B2 was qualitatively inhibited. The biosynthesis of ergosterol was significantly inhibited by the application of LEO, and morphological alterations and damage to the integrity of the fungal cell membrane were observed. The LEO has a high potential for use as an antifungal and antimycotoxigenic agent against A. carbonarius, A. flavus and A. ochraceus

Fungi and their metabolites in grain from individual households in Croatia

ABSTRACT A total of 117 fungal metabolites were detected in grains collected in Gunja-G (flooded village) and Gornji Stupnik-GS (control village), located in the Zagreb County, Croatia. Major mycotoxins and derivatives (17), ergot alkaloids (14), Fusarium (23), Aspergillus (18), Penicillium (18), Alternaria (7) and other fungal and unspecific metabolites (20) were found. A higher number of metabolites co-occurred per sample in grains from G (115) than in GS (91). Regulated mycotoxins were below maximum limits except fumonisins B1,2 in 15–20% of grains and aflatoxin B1. Fusarium metabolites contaminated more than 50% of grains at both locations. Besides FB1,2, bikaverin, aurofusarin, culmorin and 15-hidroxyculmorin were detected at relatively high concentrations. Ergot alkaloids were detected at 2–18 times higher concentrations in grains from G as compared to GS. Majority of Aspergillus mycotoxins were present at a low frequency (5–15%). Penicillium metabolites recovered with higher frequency in GS (55–70%) than in G (20–55%). Alteranaria metabolites prevailed in grains from G (60–80%).

Improved and adopted murine models to combat pulmonary aspergillosis.

The insufficient basic and clinical knowledge about invasive mold infections necessitated to review aspergillosis rodent models. The scope of this review has two major aspects. (1) It briefly summarizes Aspergillus toxicoses, the adverse effects of Aspergillus mycotoxins, the virulence factors of Aspergillus fumigatus, and how mild Aspergillus infections can turn to immunosuppressive diseases, ultimately to lethal invasive pulmonary aspergillosis. (2) The second major aspect of the review deals with earlier and recent murine models of pulmonary aspergillosis. Particular attention will be paid to the development of unified and generally applicable methods to detect, follow, and combat aspergillosis by medical treatments. Additionally, the review raises the question of responsibility regarding the application of immunosuppressive agents that initiate, contribute, and aggravate aspergillosis. Future studies of immunosuppression by chemical agents impacting aspergillosis deserve more studies.

Fusarium and Aspergillus mycotoxins contaminating wheat silage for dairy cattle feeding in Uruguay.

Wheat is one of the most important cultivated cereals in Uruguay for human consumption; however, when harvest yields are low, wheat is usually used in ensiling for animal feeding. Ensiling is a forage preservation method that allows for storage during extended periods of time while maintaining nutritional values comparable to fresh pastures. Silage is vulnerable to contamination by spoilage molds and mycotoxins because ensilage materials are excellent substrates for fungal growth. The aim of the study was to identify the mycobiota composition and occurrence of aflatoxins and DON from wheat silage. A total of 220 samples of wheat were collected from four farms in the southwest region of Uruguay were silage practices are developed. The main fungi isolated were Fusarium (43%) and Aspergillus (36%), with Fusarium graminearum sensu lato and Aspergillus section Flavi being the most prevalent species. Aflatoxin concentrations in silo bags ranged from 6.1 to 23.3 μg/kg, whereas DON levels ranged between 3000 μg/kg and 12,400 μg/kg. When evaluating aflatoxigenic capacity, 27.5% of Aspergillus section Flavi strains produced AFB1, 5% AFB2, 10% AFG1 and 17.5% AFG2. All isolates of F. graminearum sensu lato produced DON and 15-AcDON. The results from this study contribute to the knowledge of mycobiota and mycotoxins present in wheat silage.

Effects of cropping systems on food and feed contamination by Fusarium and Aspergillus mycotoxins

Certaines especes de champignons des genres Fusarium et Aspergillus sont a l'origine des principales contaminations au champ des denrees agricoles par des mycotoxines. La prevention des consequences toxicologiques de ces contaminations passe actuellement par une adaptation des systemes de culture. Compte tenu des manques importants de connaissances relatives aux facteurs declenchant la production des toxines, c'est surtout sur la prevention des risques d'infection par les especes toxinogenes que se fondent les strategies. Sur Fusarium comme sur Aspergillus, les techniques culturales permettant d' empecher la survie de l'inoculum sont un levier essentiel, notamment le choix de semences non contaminees et le raisonnement de la succession des cultures, auxquels il faut ajouter pour Fusarium le role primordial de la gestion des residus de cultures. Le choix d'une date de semis adaptee de la culture permet pour les deux genres de mettre en oeuvre des strategies d'evitement, et une baisse de la densite de semis permet une attenuation des attaques en culture. En revanche, les attenuations par pilotage de l'alimentation azotee et hydrique sont beaucoup plus difficiles a realiser, compte tenu des differences de reaction des mecanismes d'infection des pathosystemes aux etats nutritionnels des plantes. De meme, la protection fongicide est a considerer au cas par cas, compte tenu de ses effets variables sur les communautes fongiques presentes sur les cultures. Enfin, la selection varietale de cultures resistantes a l'accumulation de mycotoxines a rencontre plus de succes pour Fusarium, notamment chez les cereales et surtout le ble, que pour Aspergillus. Toutes ces techniques de prevention ne prennent un sens que dans des ensembles coherents de pratiques agricoles, dont les effets sur l'accumulation de mycotoxines en interaction avec le climat sont encore mal connus compte tenu du manque relatif de travaux menes a l'echelle du systeme de culture. Compares aux systemes conventionnels, les systemes en agriculture biologique semblent neanmoins, a ce stade des connaissances, moins ou similairement contamines.

Analytical method for the determination of sterigmatocystin in grains using high-performance liquid chromatography–tandem mass spectrometry with electrospray positive ionization

A sensitive LC–electrospray positive ionization (ESI+) MS/MS method was developed and validated for the determination of the Aspergillus mycotoxin, sterigmatocystin, in grains at levels down to less than 0.5 μg/kg. Recovery was found to be greater than 97% for wheat grains.

Secondary metabolite profiling, growth profiles and other tools for species recognition and important Aspergillus mycotoxins

Species in the genus Aspergillus have been classified primarily based on morphological features. Sequencing of house-hold genes has also been used in Aspergillus taxonomy and phylogeny, while extrolites and physiological features have been used less frequently. Three independent ways of classifying and identifying aspergilli appear to be applicable: Morphology combined with physiology and nutritional features, secondary metabolite profiling and DNA sequencing. These three ways of identifying Aspergillus species often point to the same species. This consensus approach can be used initially, but if consensus is achieved it is recommended to combine at least two of these independent ways of characterising aspergilli in a polyphasic taxonomy. The chemical combination of secondary metabolites and DNA sequence features has not been explored in taxonomy yet, however. Examples of these different taxonomic approaches will be given for Aspergillus section Nigri.

Development of a liquid chromatography/time‐of‐flight mass spectrometric method for the simultaneous determination of trichothecenes, zearalenone and aflatoxins in foodstuffs

A liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) method based on time-of-flight MS (TOFMS) with a real-time reference mass correction technique was developed for the simultaneous determination of Fusarium mycotoxins (nivalenol, deoxynivalenol, fusarenon X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, HT-2 toxin, T-2 toxin, diacetoxyscirpenol, zearalenone) and Aspergillus mycotoxins (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2) in corn, wheat, cornflakes and biscuits. Samples were cleaned up with a MultiSep #226 column. Detection of the mycotoxins was carried out in exact mass chromatograms with a mass window of 0.03 Th. Calibration curves were linear from 2 to 200 ng x mL(-1) for trichothecenes and zearalenone, and 0.2 to 20 ng x mL(-1) for aflatoxins, by 20 microL injection. The limits of detection ranged from 0.1 to 6.1 ng x g(-1) in foodstuffs analyzed in this study. The LC/TOFMS method was found to be suitable for the screening of multiple mycotoxins in foodstuffs rapidly and with high sensitivity, and its performance was demonstrated for the confirmation for target mycotoxins.

An isolate of Aspergillus flavus used to reduce aflatoxin contamination in cottonseed has a defective polyketide synthase gene.

Contamination of certain foods and feeds with the highly toxic and carcinogenic family of Aspergillus mycotoxins, the aflatoxins, can place a severe economic burden on farmers. As one strategy to reduce aflatoxin contamination, the non-aflatoxin-producing A. flavus isolate AF36 is currently being applied to agricultural fields to competitively exclude aflatoxin-producing Aspergillus species. We now show that the polyketide synthase gene (pksA) required for aflatoxin biosynthesis in AF36, and in other members of the same vegetative compatibility group, possesses a nucleotide polymorphism near the beginning of the coding sequence. This nucleotide change introduces a premature stop codon into the coding sequence, thereby preventing enzyme production and aflatoxin accumulation.

Comparative study of Aspergillus mycotoxin production on enriched media and construction material

Isolates of Aspergillus flavus and Aspergillus fumigatus from indoor air were compared with a known mycotoxin producer for their capacity to produce mycotoxins on a variety of enrichment media and with growth on indoor substrates such as ceiling tile and wall board. In enrichment media, four of seven isolates of A. flavus produced at least one aflatoxin and both isolates of A. fumigatus produced mycotoxins. The spectrum of mycotoxins and their concentrations varied with the strain and medium. When the mycotoxin-positive strains were grown to a dense concentration on indoor construction and finishing materials such as ceiling tile and wall boards, mycotoxins were not detected in extracts of the materials. Colonization of indoor surfaces by mycotoxin-producing strains of A. flavus and A. fumigatus may not necessarily expose inhabitants to mycotoxins or result in production of mycotoxins.

Peniciluum and Aspergillus Mycotoxins - Diagnostic Characters and Quantitative Data from Commodities and Cultures

Peniciluum and Aspergillus Mycotoxins - Diagnostic Characters and Quantitative Data from Commodities and Cultures